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Caltech Authors (California Institute of Technology)
Article . 1997
Full-Text: https://doi.org/10.1093/nar/25.6.1307
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Nucleic Acids Research
Article . 1997 . Peer-reviewed
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Nucleic Acids Research
Article
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Nucleic Acids Research
Article . 1997
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https://dx.doi.org/10.1093/nar...
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Optimization of DNA shuffling for high fidelity recombination

descriptionPublicationkeyboard_double_arrow_right Article 15 Mar 1997 United States Publisher:Oxford University Press (OUP)Journal:Nucleic Acids Research, volume 25, pages 1,307-1,308 (eissn: 1362-4962, Copyright policy )
Authors: Zhao, Huimin; Arnold, Frances H.;

doi: 10.1093/nar/25.6.1307

pmid: 9092645

pmc: PMC146579

Optimization of DNA shuffling for high fidelity recombination

Abstract

A convenient 'DNA shuffling' protocol for random recombination of homologous genes in vitro with a very low rate of associated point mutagenesis (0.05%) is described. In addition, the mutagenesis rate can be controlled over a wide range by the inclusion of Mn2+or Mg2+during DNase I digestion, by choice of DNA polymerase used during gene reassembly as well as how the genes are prepared for shuffling (PCR amplification versus restriction enzyme digestion of plasmid DNA). These protocols should be useful for in vitro protein evolution, for DNA based computing and for structure-function studies of evolutionarily related genes.

Country
United States
Related Organizations
Keywords

Recombination, Genetic, 570, Manganese, DNA, in-vitro, Biological Evolution, Polymerase Chain Reaction, Recombinant Proteins, fragmentation, evolution, Mutagenesis, Site-Directed, Deoxyribonuclease I, Point Mutation, Indicators and Reagents, Magnesium, Subtilisins, polymerases, Gene Library, Plasmids

  • BIP!
    Impact byBIP!
    selected citations
    These citations are derived from selected sources.
    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    		 190
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    		 Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    		 Top 1%
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    		 Top 1%
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
190
Top 10%
Top 1%
Top 1%
Green
gold